Quantifying Host-Parasite Interactions

Host cell and intracellular T. cruzi amastigote numbers were assessed as described with minor modifications [34 (link)]. Briefly, NHDF were plated at 1.5 x 10³ per well in 384 well plates and infected with MOI 1.25 for 2 hours before incubation in phenol-free media at the indicated glucose concentration. At 18 hpi, the indicated concentration of 2-DG was added, and at 66 hpi media was removed and host cell number and parasite number were assessed using 10 μL of CellTiter-Fluor (Promega) and 10 μL of Beta-Glo (Promega) per well, respectively.

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Shah-Simpson S., Lentini G., Dumoulin P.C, & Burleigh B.A. (2017). Modulation of host central carbon metabolism and in situ glucose uptake by intracellular Trypanosoma cruzi amastigotes. PLoS Pathogens, 13(11), e1006747.

Publication 2017

CellTiter-Fluor Beta-Glo

Cell Glucose Intracellular Parasite Phenol Promega

Corresponding Organization : Harvard University

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Variable analysis

independent variables

Glucose concentration
2-DG concentration

dependent variables

Host cell number
Intracellular T. cruzi amastigote number

control variables

Cell type (NHDF)
Infection MOI (1.25)
Infection duration (2 hours)
Incubation time (18 hpi, 66 hpi)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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